- 作者：Zheng Zhao ; PhD&lowast ; ; &dagger ; ; Hongchen Liu ; PhD&lowast ; ; zhaozheng6912@sohu.com" class="auth_mail" title="E-mail the corresponding author ; Dongsheng Wang ; PhD&lowast ;
- 关键词：ADAM28 ; apoptosis ; differentiation ; human dental pulp stem cells ; proliferation
- 刊名：Journal of Endodontics
- 出版年：2011
- 出版时间：March 2011
- 年：2011
- 卷：37
- 期：3
- 页码：332-339
- 全文大小：897 K
Methods
After ADAM28 eukaryotic plasmid and antisense oligodeoxynucleotides (AS-ODNs) were constructed and respectively transfected into HDPSCs by Lipofectamine 2000, the ADAM28 expression levels among diverse groups were estimated by reverse transcription polymerase chain reaction (RT-PCR) and western blotting. Methabenzthiazuron (MTT) and cell cycle assays were used to test the HDPSCs proliferation activity. Annexin V- fluorescein isothiocyanate (FITC)/propidium iodide and alkaline phosphatase analysis were performed respectively to measure apoptosis and the cytodifferentiation level. Immunocytochemistry and western blotting were performed to determine the effects of ADAM28 eukaryotic plasmid on HDPSCs expressing dentin sialophosphoprotein (DSPP), dentin matrix protein 1, and bone sialoprotein.
Results
ADAM28 could be correctly transcribed, translated, and expressed in HDPSCs. The ADAM28 AS-ODN group displayed the highest optical density value, whereas the eukaryotic plasmid group showed the lowest, which suggested that ADAM28 had a negative regulatory effect on the proliferation of HDPSCs. ADAM28 eukaryotic plasmid could significantly inhibit the HDPSC proliferation, promote specific differentiation of HDPSCs, induce apoptosis, and enhance the DSPP expression, whereas ADAM28 AS-ODN produced the opposite effects.
Conclusions
Our results proved that ADAM28 might actively participate in manipulating the proliferation, differentiation, and apoptosis of HDPSCs.